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减体积肝移植模型大鼠肝脏能量差异蛋白的表达
作者姓名:刘 静  李 立  冉江华  张升宁  李来邦  张熙冰  高 扬  陈奕明
作者单位:昆明医科大学附属甘美医院、昆明市第一人民医院肝胆胰外科,云南省昆明市 650011
摘    要:背景:目前,蛋白质组学是一项比较成熟的科研技术,已经在肝移植基础研究领域中初步得到应用,但在大鼠减体积肝移植相关研究中的应用未曾报道。 目的:应用蛋白质组学相关技术探讨大鼠减体积肝移植后与肝脏能量代谢蛋白相关的差异蛋白。 方法:肝移植的供体是Lewis雌性大鼠,受体是Wistar雄性大鼠,供受体体质量差约20 g;移植肝质量/受体肝质量≈50 %。采用改良减体积肝移植模型。获取供体肝组织过程中按照要求进行减体积,修整供体肝脏,将门静脉和肝下下腔静脉分别套入不同型号的套管以备套入受体的门静脉和肝下下腔静脉,胆道用细小支撑管植入供体的胆管中以备插入到受体的胆管中;最后进行受体的肝移植,受体先切除受体的原来肝脏组织,然后将准备好的供体肝脏植入受体中。造模后1,3和7 d获取移植肝脏组织,然后与预先获取冻存的供、受体肝脏组织应用蛋白组学技术建立双向凝胶电泳图谱,再利用串联质谱分析及数据库对差异表达的蛋白质点进行鉴定。 结果与结论:采用变化倍数大于10倍为标准进行差异蛋白点的选择,结果总共发现了72个差异点,通过鉴定,最终32个蛋白的功能比较明确,其中有ATP合成酶β亚基、电子转化黄素蛋白β多肽和质子转运ATP合成酶3个蛋白参与了细胞能量代谢的过程,其分布于肝移植后的第1和7天,占6%。 中国组织工程研究杂志出版内容重点:肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接:

关 键 词:实验动物  消化系统损伤模型  减体积肝移植  肝脏  蛋白质组学  串联质谱分析  差异蛋白  能量蛋白  ATP合成酶  
收稿时间:2015-02-03

Expression of hepatic energy proteins following reduced-size liver transplantation in rats
Authors:Liu Jing  Li Li  Ran Jiang-hua  Zhang Sheng-ning  Li Lai-bang  Zhang Xi-bing  Gao Yang  Chen Yi-ming
Institution:Department of Hepato-biliary-pancreatic Surgery, the First People’s Hospital of Kunming and the Ganmei Affiliated Hospital of Kunming Medical University, Kunming 650011, Yunnan Province, China
Abstract:BACKGROUND: At present, the proteome is a mature technology that has been applied in basic research fields related to liver transplantation. But, it has been not reported in research related to reduced-size liver transplantation. OBJECTIVE: To explore the expression of differential proteins related to hepatic energy metabolism following reduce-size liver transplantation in rats by using by proteomic technology. METHODS: The improved model of reduced-size liver transplantation was used in this experiment. The donor was health female Lewis rats and the recipient was male Wistar rats for liver transplantation. The difference between the donor and the recipient was about 20 g. The weight of donor liver/the weight of recipient donor was approximately equal to 50%. The donor liver tissue was harvested and trimmed to the required size. The portal vein and infrahepatic vena cava were cannulated, and the biliary tract was implanted into the donor bile duct for  transplantation. Then the donor was transplanted into the recipient after the removal of original liver tissue. Hepatic specimens were harvested by 1, 3 and 7 days after reduced-size liver transplantation. Then, the harvested specimens were compared with the normal donor and recipient liver tissue that were previously harvested and frozen, to generate two-dimensional gel electrophoresis profile using proteome technology. Then tandem mass spectrometry and databases analysis were performed after two-dimensional electrophoresis for identifying differential protein stains. RESULTS AND CONCLUSION: In this experiment, 72 differential protein stains with over lo-fold changes were selected. After identification, 32 proteins showed clear functions, and among them three differential proteins (ATP synthase beta subunit, electron-transferring flavoprotein beta peptide and proton-transferring ATP synthase) were involved in the process of cell energy metabolism. The proteins were distributed on 1 and 7 days after reduce-size liver transplantation, accounting for 6%.
Keywords:Liver Transplantation  Proteomics  ATP Synthase Complex  Energy Metabolism  
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